Upload
others
View
1
Download
0
Embed Size (px)
Citation preview
2
3
The 4th Leading Special Lecture
New Aspect in Clinical Chemistry
What is the role of "Analytical Chemistry" in Veterinary Medicine?
March 13, 2013, 16:00~17:30
Lecture Hall, Graduate School of Veterinary Medicine,
Hokkaido University, JAPAN
Program
Chairpeson: Prof. Mitsuyoshi Takiguchi (Hokkaido Univ, JAPAN)
16:00~16:20
Dr. Kei Nomiyama (Ehime Univ, JAPAN)
Organohalogen compounds and their metabolites in the blood of pet dogs and cats, and in pet food
16:20~16:30
Prof. Mitsuyoshi Takiguchi (Hokkaido Univ, JAPAN)
Future expectation for applications of metabolomics
in clinical veterinary medicine
16:30~17:30
Prof. Robert H Weiss (UC Davis, USA)
Metabolomics in Kidney Cancer: From biofluids to new therapeutic targets.
4
Dr. Kei Nomiyama, Ph.D.
Senior Assistant Professor of Environmental Chemistry and Ecotoxicology
division of the Center for Marine Environmental Studies (CMES), Ehime
University
1999--2003, Faculty of Environmental Symbiotic Sciences Prefectural
University of Kumamoto, Japan;
2003--2005, Laboratory of Environmental Water Sciences, Graduate School of Environmental Symbiotic
Sciences, Prefectural University of Kumamoto, Japan,
Received Masters Degree in Environmental Symbiotic Sciences;
2005--2008, Laboratory of Environmental Water Sciences, Graduate School of Environmental Symbiotic
Sciences, Prefectural University of Kumamoto, Japan, Received Ph.D. Environmental Symbiotic Sciences;
2007--2008, Research Fellowship for Young Scientist (DC2) of Japan Society for the Promotion of Science
(JSPS);
2008--2011, Assistant Professor, CMES, Ehime University;
2011-present Senior Assistant Professor, CMES, Ehime University
5
Organohalogen compounds and their metabolites
in the blood of pet dogs and cats, and in pet food
Kei Nomiyama1*, Hazuki Mizukawa1, Susumu Nakatsu2, Hisato Iwata1, Akira Kubota3, Miyuki
Yamamoto1, Jean Yoo1 and Shinsuke Tanabe1 1 Center for Marine Environmental Studies (CMES), Ehime University, Japan.
2 Nakatsu Veterinary Surgery, Osaka, Japan. 3 Biology Department, Woods Hole Oceanographic Institution, U.S.A.
* E-mail: [email protected]
Polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) have been widely
used for industrial applications since the 1960s and 1970s. As a consequence, PCB and PBDE levels have
been increasing dramatically in the environment, wildlife and humans, because of their persistent properties
and bioaccumulation characteristics. Toxic effects of these compounds on endocrine systems and
neurodevelopment are already well known. It is suspected that the thyroid hormone homeostasis is
disturbed by not only PCBs and PBDEs but also their hydroxylated metabolites. Hydroxylated PCBs (OH-
PCBs) are formed by the oxidative metabolism of PCBs by cytochrome P450 (CYP) monooxygenase
enzyme systems. Hydroxylated PBDEs (OH-PBDEs) are well-known metabolites of PBDEs, and also natural
substances found in marine organisms.
Recently, a study found higher concentrations of PBDEs in pet cats than in human serum, and it has
been hypothesized that increases of feline hyperthyroidism (HT) are related to increased PBDE exposure,
with key routes of exposure being diet and ingestion of house dust. Moreover, increased thyroid
hyperfunction disease may be linked to OH-PCBs and OH-PBDEs derived from metabolites of PCBs and
PBDEs, diet, natural products, and/or demethylation of MeO-PBDEs. However, information on status of
hydroxylated metabolites of PCBs and PBDEs in pet animals and pet food are limited.
The present study determined the concentrations and accumulation patterns of organohalogen
contaminants such as PCBs and PBDEs and their metabolites (OH-PCBs, OH-PBDEs and MeO-PBDEs) in
the whole blood of pet cats and pet dogs collected from veterinary hospital in Japan. Moreover, to estimate
the dietary PCB and PBDE exposure levels and biotransformation to hydroxylated metabolites in pet
animals, PCB, PBDE and their derivatives (OH-PCBs, OH-PBDEs, and MeO-PBDEs) in the pet foods were
also analyzed.
Concentrations of OH-PCBs in the blood of dogs and cats were 220 ± 260 pg/g and 150 ± 80
pg/g, respectively. Tri- and penta-chlorinated OH-PCB congeners were predominant in cat blood. In
6
contrast,accumulation of hexa- through octa-chlorinated OH-PCBs has been found in dog blood.
These results were similar to pattern of previous terrestrial mammal study. In contrast, OH-PCB
levels in pet food (range: 0.52-1.3 pg/g) were negligible. These results indicate that the OH-PCBs
detected in pet dogs and cats are metabolites of PCBs because pet food did not contain OH-PCBs.
PBDE levels in pet bloods were similar to those in dry pet food, and BDE209 was the dominant
congeners in both bloods and food. The residual levels of 6OH-BDE47 and 2′MeO-BDE68 in the
blood of pet cats were similar. MeO-PBDEs levels in seafood rich (wet) cat diet were one order
higher than those in cat bloods, while OH-PBDEs levels in cat food were one to two orders lower
than the cat bloods. Among the OH-/MeO-PBDE isomers, only 6OH-/MeO-BDE47 and 2′OH-/MeO-
BDE68 were detected in the blood of dogs and cats and pet food. These two abundant isomers are
produced naturally by marine organisms. These results suggested that pet cats are exposed to
MeO-PBDEs through cat food such as fish flavor. On the other hand, low concentrations of OH-
/MeO-PBDEs in dog blood and dog food were found. This result suggests that uptake of from
naturally produced OH-/MeO-PBDEs within the dog foods are very low. When in vitro demethylation
experiment of MeO-PBDEs using dogs and cats liver microsomes was conducted, the demethylation
of MeO-PBDEs in dogs and cats microsomes have been confirmed. This result shows large
percentage of OH-PBDEs found in the cat blood might be direct intake from food and
biotransformation of MeO-PBDEs. On the other hand, it can be suggested that dogs may have high
metabolic capacities for OH-/MeO-PBDEs through phase II reaction, or low TTR binding potencies
of OH-PBDEs. These results suggested that induction of thyroid dysfunction in pet cats (which are
originally carnivorous) might be due to forced fish eating through seafood.
Metabolic capacities of dogs and cats were compared by the in vitro metabolism test of PCBs
using hepatic microsomes. After exposure to 62 PCB mixtures, 4’OH-CB18, 3’OH-CB28, 4’OH-CB79, 4OH-
CB107, and 13 unknown peaks (3-5Cl OH-PCBs) were found in cat liver microsomes. The homolog pattern
identified in this experiment was similar to those in the pet cat blood. In contrast, 13 identified OH-PCBs and
22 unknown OH-PCBs (3-8Cl OH-PCB) were detected in the dog liver microsomes, after exposed to PCB
mixtures. Different homolog pattern between this experiment and pet dog blood was observed. It suggests
lower chlorinated OH-PCBs were more easily eliminated through phase II conjugation reaction in dog body.
These results indicate possibility of induction of adverse health effects, especially feline HT, due to
the consumption of cat feed made from fishery product and specific metabolic capacities. In future studies,
we need to investigate the metabolic capacities to halogenated compounds including phase II conjugation
reactions, and assess the toxicological risk posed by these hydroxylated metabolites.
7
8
Prof. Mitsuyoshi Takiguchi
Professor of Laboratory of Veterinary Internal Medicine Graduate School of Veterinary Medicine, Hokkaido University EDUCATION: April 1983 - March 1987 Bachelor of Veterinary Science Degree Faculty of Veterinary Medicine, Hokkaido University April 1987 - March 1989 Master of Veterinary Medical Science Degree Graduate School of Veterinary Medicine, Hokkaido University December 2000 PhD Degree Graduate School of Veterinary Medicine, Hokkaido University PREVIOUS EMPLOYMENT: 1989 Staff Veterinarian Yokohama Animal Medical Center 1992 Visiting Researcher Department of Veterinary Clinical Science, College of Veterinary Medicine The Ohio State University 1993 Assistant professor Department of Veterinary Clinical Sciences, Faculty of Veterinary Medicine Hokkaido University 2000 Lecturer Department of Veterinary Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University
2004 Associate professor Department of Veterinary Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University
2004 Professor Department of Companion Animal Sciences, School of Veterinary Medicine, Rakuno Gakuen University
2007 Professor Department of Veterinary Clinical Sciences, Graduate School of Veterinary Medicine Hokkaido University
9
Future expectation for applications of metabolomics
in clinical veterinary medicine
Mitsuyoshi Takiguchi, DVM, PhD
Laboratory of Veterinary Internal Medicine
Graduate School of Veterinary Medicine, Hokkaido University
The field of metabolomics continues to grow rapidly over the last decade and has been proven to be a
powerful technology in predicting and explaining complex phenotypes in diverse biological systems.
Application of metabolomics has been increasing in the medical field. In this short talk, I would like to
talk about future expectation for applications of metabolomics in clinical veterinary medicine.
10
Prof. Robert H. Weiss, MD Professor of Medicine, University of California, Davis, School of Medicine
Chief of Nephrology, VA Medical Center, Sacramento, CA
Member, Cancer Center, UC Davis
EDUCATION
College: University of California, Santa Cruz. B.A. Chemistry, 1976
Graduate School: UCLA M.S. Physical Chemistry, 1978
M.S. Medical Physics, 1980
Medical School: University of California, Irvine M.D. Medicine, 1984
Residency: California Pacific Medical Center Internal Medicine, 1987
Fellowship: University of California, San Francisco Nephrology, 1991
HONORS/AWARDS/EXTRAMURAL
1972 University of California President's Scholarship
1974 National Science Foundation Undergraduate Research Award
1976 University of California, Santa Cruz, College Honors
1978 NIH National Research Service Award
1983 American Cancer Society Medical Student Fellowship
1995 Elected to Fellowship, American College of Physicians
1997 National Kidney Foundation Clinical Scientist Award
2001 Nominated for Dean’s Mentoring Award
2002 Nominated for Dean’s Mentoring Award
2003 Nominated for Dean’s Mentoring Award
2004 Nominated for Dean’s Mentoring Award
2005 Joan Oettinger Award for Outstanding Cancer Research
2004 VA Research Honors
2004 VA Medical Research Technologies Researcher spotlight
2006-pres Associate Member, NCI Early Detection Research Network (EDRN)
PROFESSIONAL POSITIONS AND APPOINTMENTS:
1991-present Staff Nephrologist, VA Northern California System of Clinics
1991-present Member, Physiology Graduate Group
1991-1998 Assistant Professor of Medicine, University of California, Davis, School of Medicine
11
1998-2003 Associate Professor of Medicine, University of California, Davis, School of Medicine
2000-present Member, Cell and Developmental Biology Graduate Group, UCD
2002-present Member, UC Davis Cancer Center
2002-present Member, Biochemistry and Molecular Biology Graduate Group, UCD
2003-present Professor of Medicine, University of California, Davis, School of Medicine
2003-present Member, Immunology Graduate Group, UCD
2004-present Member, Western Association of Physicians
2004-present Chief, Nephrology Section, Mather (Sacramento) VAMC
2006-present Member, Comparative Pathology Graduate Group
2009-present Editor-in-Chief, Cancers http://www.mdpi.com/journal/cancers
RECENT PUBLICATIONS (2011~)
Chen H, Li C, Huang J, Cung T, Seiss K, Beamon J, Carrington MF, Porter, LC, Burke PS, Yang Y, Ryan BJ,
Liu R, Weiss RH, Pereyra F, Cress WD, Brass AL, Rosenberg ES, Walker BD, Yu XG, Lichterfeld M.
CD4+ T cells from elite controllers resist HIV-1 infection by selective upregulation of p21 (cip-1/waf-1).
Journal of Clinical Investigation. Apr 1;121(4):1549-60, 2011.
Weiss RH, Inoue H. Diagnosis and Treatment of Kidney Cancer. Nephrology CyberRounds., 2011
http://www.cyberounds.com/cmecontent/04/87/art487.html?preview=on
Kim, K., Taylor, S.L., Ganti, S., Guo, L., Osier, M., Weiss, R.H. Urine metabolomic analysis identifies
potential biomarkers and pathogenic pathways in kidney cancer, Omics, May;15(5):293-303, 2011.
Ganti., S., Weiss, R.H. Urine metabolomics for kidney cancer detection and biomarker discovery. Urologic
Oncology: Seminars and Original Investigations, 2011 Sep-Oct;29(5):551-7. 2011
Buzon, M., Seiss, K., Weiss, R.H., Brass, A., Rosenberg, E., Pereyra, F., Yu, X., Lichterfeld, M. Inhibition of
HIV-1 integration in ex vivo infected CD4 T cells from elite controllers, Journal of Virology, 2011
Sep;85(18):9646-50. 2011
Inoue, H., Hwang, S.H., Wecksler, A.T., Hammock, B.D., Weiss, R.H. Sorafenib attenuates p21 in kidney
cancer cells and augments cell death in combination with DNA-damaging chemotherapy, Cancer
Biology & Therapy, 2011 Nov 1;12(9):827-36, 2011
Ganti, S., Taylor, S.L., Kim, K., Hoppel, C.L., Guo, L., Yang., J., Evans, C., Weiss, R.H. Urine acylcarnitines
are altered in human kidney cancer, International Journal of Cancer, Jul 5, 2011
Weiss R.H, Kim K. Metabolomics in the Study of Kidney Diseases. Nature Reviews Nephrology. Oct
25;8(1):22-33, 2011
Ganti, S., Taylor,S.L., Aboud,O.A., Yang,J., Evans,C., Osier,M.V., Alexander,D.C., Kim,K., Weiss,R.H.
Simultaneous multiple matrix metabolomics analysis of a mouse kidney cancer xenograft yields
potential tumor biomarkers. Cancer Research, Jul 15;72(14):3471-9, 2012
12
Zivkovic, A.M., Yang, J., Georgi, K., Hegedus, C., Nording, M.L., O’Sullivan, A., German, J.B., Hogg, R.J.,
Weiss, R.H., Bay, C., Hammock, B.D. Serum oxylipin profiles in IgA nephropathy patients reflect
kidney functional alterations. Metabolomics, December 2012, Volume 8, Issue 6, pp 1102-1113
Hoffman, M.D., Stuempfle, K.J., Fogard, K., Hew-Butler, T., Winger, J., Weiss, R.H. Urine Dipstick Analysis
for Identification of Runners Susceptible to Acute Kidney Injury following an Ultramarathon, Journal of
Sports Sciences, 2012 Oct 4. [Epub ahead of print]
Inoue, H., Kauffman, M., Shacham, S., Landesman, Y., Weiss, R.H. Inhibition of nuclear export by CRM1
attenuation causes apoptosis in kidney cancer cells. Journal of Urology, 2012 Oct 15. S0022-
5347(12)05217-2. [Epub ahead of print]
Abu Aboud, O., Weiss, R.H. New opportunites from the cancer metabolome. Clinical Chemistry, 2013
Jan;59(1):138-46.
Wettersten, H., Hwang, S.H., Li, C., Shiu, E.Y., Wecksler, A.T., Hammock, B.D., Weiss, R.H. A novel p21
attenuator which is structurally related to sorafenib, Cancer Biology & Therapy, 2013 Jan 8;14(3).
Ulu, A., Harris, T.R., Morisseau, C., Miyabe, C., Inoue, H., Schuster, G., Dong, H., Iosef, A.-M., Weiss, R.H.,
Chamvimonvat,. N., Imig, J.D., Hammock, B.D. Anti-inflammatory Effects of Omega-3 Polyunsaturated
Fatty Acids and Soluble Epoxide Hydrolase Inhibitors in Angiotensin-II Dependent Hypertension.
Journal of Cardiovasular Pharmacology, in press
Liu, R., Wettersten, H., Park, S.-H., Weiss, R.H. Small-molecule inhibitors of p21 as novel therapeutics for
chemotherapy-resistant kidney cancer, Future Medicinal Chemistry, in press
Martin KS, Soldi C, Candee KN, Wettersten HI, Weiss RH, Shaw JT. From bead to flask: Synthesis of a
complex β-amido-amide for probe-development studies. Beilstein J Org Chem. 2013;9:260-4
Wettersten, H., Weiss, R.H. Potential biofluid markers and targets for clear cell renal carcinoma diagnosis
and treatment. Nature Reviews Urology (invited review), in revision.
Kim, K., Mall, C., Taylor, S.L., Hitchcock, S., Zhang, C., Jones, A.D., Chapman, A., Weiss, R.H. Mealtime,
temporal, and daily variability of the human urinary and plasma metabolomes in a tightly controlled
environment. J. Proteome Research, in revision
Zhang, G., Panigrahy, D., Mahakian, L., Yang, J., Liu, J.-Y., Lee, K.S.S., Wettersten, H., Ulu, A., Hu, X.,
Tam, S., Hwang, S.H., Ingham, E., Weiss, R.H., Ferrara, K.W., Hammock, B.D. Epoxy metabolites of
docosahexaenoic acid (DHA) inhibit angiogenesis, tumor growth and metastasis. Proceedings of the
National Academy of Sciences (USA), in revision.
13
Metabolomics in Kidney Cancer:
From biofluids to new therapeutic targets
Robert H. Weiss, MD
University of California, Davis
Of the omics sciences, metabolomics is one of the newer methodologies. In this technique, all of the
small molecule substances produced during bodily processes are measured. This includes endogenous
metabolites as well as small peptides, amino acids, and (usually) body flora and drug metabolites. Rather
than being participators in these processes, as are genes and proteins, the metabolites produced are
sentinels of “what is actually happening” in an organism. The so-called “cancer metabolome” is loosely
defined as the entire suite of relatively low molecular weight (~<1500 daltons) metabolites germane to
cancer and includes their changes relative to a control non-cancerous group of patients or tissues1. Using
this data, metabolic pathways altered in a specific cancer can be identified, and once a metabolic pathway is
determined to be altered in a specific tumor or in the systemic response to it, and a full validation (in vitro
and/or in vivo) of the role of this metabolite takes place, pharmacologic inhibitors or activators of such
pathways can be either designed or re-purposed to be used as potential new chemotherapeutics2.
Most of the metabolic processes in the body, such as those involving energetics and amino acid
catabolism, are common to all living cells. However, it is logical that in cells which are highly proliferative or
which have deranged apoptosis, such as cancer cells, there would be at least some biochemical pathways
that are enhanced or diminished. For example, in order for a cell to grow rapidly, there exists the necessity
for high energy requirements as well as for the provision of membrane and other cellular building blocks. In
addition, there are likely to be profound changes in metabolic pathways such as glycolysis (e.g. the Warburg
effect), amino acid metabolism, and fatty acid oxidation. Due to its appetite for transforming many metabolic
pathways to its own advantage (which ultimately leads to its causing mortality), cancer is ideally suited for
metabolomics analysis. Since cancer, especially in the later stages, so profoundly usurps normal metabolic
processes, it can be considered the ultimate metabolic disease.
As applied to RCC, metabolomics evaluations can be performed on urine, blood, and tissue to
identify altered metabolic pathways and thereby point to new targets for therapeutic intervention3. In addition,
such signatures, if they are found to be specific to the cancer in question, can be used either singly or as
multiplexed as biomarkers. Even though it is likely that most RCCs are “walled off” from the urinary space,
the use of urinary metabolite determination has been especially fruitful and has yielded signatures of
14
tryptophan metabolism as well as energetic derangements. Similar analyses could potentially be
accomplished with cyst fluid. While to date there have been no proven specific metabolic biomarkers for
ccRCC, this technique has yielded novel targets, such as PPARα, CPT-1, and IDO2, which can be evaluated
using available pharmaceuticals.
In summary, metabolomics is a powerful new technique which can be utilized in cancer biology to
identify both markers and new therapeutic targets, and in the form of pharmacometabolomics4, can lead to
the repurposing of heretofore discarded drugs and thus lead to therapeutic advances.
References
1. Ganti, S. and Weiss, R. H.: Urine metabolomics for kidney cancer detection and biomarker discovery.
Urol Oncol, 29: 551, 2011.
2. Ganti, S., Taylor, S. L., Abu, A. O., Yang, J., Evans, C., Osier, M. V. et al.: Kidney tumor biomarkers
revealed by simultaneous multiple matrix metabolomics analysis. Cancer Res, 72: 3471, 2012.
3. Kim, K., Taylor, S. L., Ganti, S., Guo, L., Osier, M. V., and Weiss, R. H.: Urine metabolomic analysis
identifies potential biomarkers and pathogenic pathways in kidney cancer. OMICS, 15: 293,
2011.
4. Weiss, R. H. and Kim, K.: Metabolomics in the study of kidney diseases. Nat Rev Nephrology, 8: 22,
2011.
15
Graduate School of Veterinary Medicine, Hokkaido University
Kita18, Nishi9, Kita-ku, Sapporo 060-0818, Japan
16